741 results about "Protein purification" patented technology

The instant invention relates to the field of protein production and purification, and in particular to compositions and processes for controlling the amount of charge variants, aggregates, and fragments of a protein of interest, as well as host cell proteins, present in purified preparations by applying particular chromatography conditions during such protein purification.

A method for purifying a polypeptide by ion exchange chromatography is described which involves changing the conductivity and/or pH of buffers in order to resolve a polypeptide of interest from one or more contaminants.

A self-cleaving element for use in bioseparations has been derived from a naturally occurring, 43 kDa protein splicing element (intein) through a combination of protein engineering and random mutagenesis. A mini-intein (18 kDa) previously engineered for reduced size had compromised activity and was therefore subjected to random mutagenesis and genetic selection. In one selection a mini-intein was isolated with restored splicing activity, while in another, a mutant was isolated with enhanced, pH-sensitive C-terminal cleavage activity. The enhanced cleavage mutant has utility in affinity fusion-based protein purification. The enhanced splicing mutant has utility in purification of proteins such as toxic proteins, for example, by inactivation with the intein in a specific region and controllable splicing. These mutants also provide new insights into the structural and functional roles of some conserved residues in protein splicing. Thus, disclosed and claimed are: a genetic system and self-cleaving inteins therefrom; bioseparations employing same; protein purification by inactivation with inteins in specific regions and controllable intein splicing; methods for determining critical, generalizable residues for varying intein activity; and products.

A method for purifying a polypeptide by ion exchange chromatography is described in which a gradient wash is used to resolve a polypeptide of interest from one or more contaminants.

The present invention relates generally to the production of recombinant human uteroglobin (rhUG) for use as a therapeutic in the treatment of inflammation and fibrotic diseases. More particularly, the invention provides processes, including broadly the steps of bacterial expression and protein purification, for the scaled-up production of rhUG according to current Good Manufacturing Practices (cGMP). The invention further provides analytical assays for evaluating the relative strength of in vivo biological activity of rhUG produced via the scaled-up cGMP processes.

An object of the present invention is to provide a novel protein having the following amino acid sequence altered for specifically and efficiently binding a protein to an immobilization carrier via the carboxy terminus. The protein is used for immobilizing a portion represented by R1-R2 on an immobilization carrier, comprising the amino acid sequence represented by the general formula R1-R2-R3-R4-R5 [wherein:

• • the sequences are oriented from the amino terminal side to the carboxy terminal side;
  • the sequence of the R1 portion is the sequence of a subject protein to be immobilized and contains neither a lysine residue nor a cysteine residue;
  • the sequence of the R2 portion may be absent, but when the sequence of the R2 portion is present, the sequence of the R2 portion is a spacer sequence composed of amino acid residues other than lysine and cysteine residues;
  • the sequence of the R3 portion is composed of two residues of amino acid represented by cysteine-X (where X denotes an amino acid residue other than lysine or cysteine);
  • the sequence of the R4 portion may be absent, but when the sequence of the R4 portion is present, the sequence of the R4 portion contains neither a lysine residue nor a cysteine residue, but contains an acidic amino acid residue capable of acidifying the isoelectric point of the entire protein comprising the amino acid sequence represented by the general formula R1-R2-R3-R4-R5; and
  • the sequence of an R5 portion is an affinity tag sequence for protein purification.

Methods of reducing high molecular weight species (HMW) formation in a sample containing a protein purified using ion exchange (IEX) chromatography are disclosed, as are a number of related methods, e.g., methods of reducing on-column denaturation of a protein in a protein sample purified using an ion exchange (IEX) column or resin. The methods share characteristics of including arginine, glycine and/or histidine in the buffers used during the ion exchange (IEX) chromatography.

The instant invention relates to modulated lysine variant species compositions comprising a protein, e.g., an antibody, or antigen-binding portion thereof, and methods, e.g., cell culture and/or protein purification methods, for producing such modulated lysine variant species compositions. Methods for using such compositions to treat a disorder, e.g., a disorder in which TNFα is detrimental, are also provided.

The present invention provides novel and improved protein purification processes which incorporate certain types of carbonaceous materials and result in effective and selective removal of certain undesirable impurities without adversely effecting the yield of the desired protein product.

Purification of recombinant proteins is performed by expressing in a host cell a fusion protein comprising: (a) a product protein domain, (b) an intein, and (c) at least one aggregator protein domain, wherein the aggregator protein domain comprises a protein that is capable of specific association with granules of polyhydroxyalkanoate (PHA).

This invention discloses one EPSPS gene enzyme immune agent case and its use method in the anti-herbicide bean, wherein, the case comprises anti-EPSPS multi-clone antigen, enzyme board with the antigen, enzyme antigens for two, gene switch bean standard product, non-transfer gene bean standard, intense wash liquid, develop liquid and terminal liquid. This invention method comprises the following steps: cloning CP4-EPSPS gene from the gene switch bean expressed in the bacillus coli; then through protein purification complex property to regroup EPSPS protein antibody immune animal to get special single clone antigen and multi-clone antigen; establishing double antigen clamp enzyme immune system test sample to determine its EPSPS protein content.

The invention provides a method for purifying protein of enzyme aggregate based on self-aggregation short-peptide induction. The method comprises the following steps of: fusing and expressing a target protein, a peptide section (preferably intein) containing a cleavage site and a short peptide with self-aggregation function into a triplet in a sequence from left to right or from right to left, forming an enzyme aggregate in an expressing process, and separating from most soluble impurities in cell lysate by adopting a centrifuging or filtering method; and then releasing the target protein into solution by inducing cutting at a cleavage site so as to achieve the purpose of purification. The method has the characteristics of low cost and simple flow, can be used for laboratory-scale high-throughput protein purification and is also beneficial to industrial-scale protein production.

The invention is directed to a method for producing a polypeptide composition comprising: combining a polypeptide with a volatile additive to form a liquid mixture and lyophilizing the liquid mixture to obtain a lyophilized polypeptide composition.

The present invention provides methods for purifying proteins. In particular, the methods employ a two-step non-affinity chromatography process without the use of an in-process tangential flow filtration step.